Institute of Fundamental Technological Research
Polish Academy of Sciences

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Dorota Lewińska

Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
Supervision of doctoral theses
1.  2019-06-11 Kupikowska-Stobba Barbara  
(IBIB PAN)
A one-step electrostatic method for encapsulation of cells in alginate-polyethersulfone microcapsules 

Recent publications
1.  Kupikowska-Stobba B., Grzeczkowicz M., Lewińska D., A one-step in vitro continuous flow assessment of protein release from core-shell polymer microcapsules designed for therapeutic protein delivery, Biocybernetics and Biomedical Engineering, ISSN: 0208-5216, DOI: 10.1016/j.bbe.2021.05.003, Vol.41, No.4, pp.1347-1364, 2021

Abstract:
Developing accurate methods for the assessment of therapeutic protein release from polymer drug delivery systems (microcapsules, microspheres, nanoparticles, 3D-printed systems) is of paramount importance for new formulation development. The most straightforward method for protein release assessment is spectrophotometric analysis of the release medium surrounding the formulation. However, direct spectrophotometric analysis is inapplicable to formulations releasing interfering compounds (co-encapsulated drugs, additives) absorbing light in the same spectrum as proteins. Conventional protein release assays also require frequent release medium sampling and replacement, which reduces their accuracy. We propose a one-step method to assess protein release from core/shell microcapsules eliminating the need for sampling and allowing selective real-time protein quantitation in the release medium. To prevent spectral interferences, released protein is differentiated from interfering compounds by employing a colorimetric protein assay reagent, forming a colour complex selectively with the protein, as the release medium. To eliminate sampling, we employed a continuous flow closed loop set-up, where the release medium is constantly circulating between microcapsule-containing tank and spectrophotometer. A series of colorimetric protein assay reagents (bromocresol green, tetrabromophenol blue, eosin B, eosin Y, biuret) were evaluated in terms of their applicability as the release medium in described system. Only biuret reagent was found compatible with proposed method due to formation of color complex stable over extended period of time and low adsorption to microcapsules. Presented method allowed effective evaluation of albumin release from alginate-polyethersulfone microcapsules with accuracy equal to conventional ‘sample and separate’ technique. Albumin release followed first-order kinetics with plateau reached after 19 h.

Keywords:
protein release, core-shell microcapsule, continuous flow apparatus, colorimetric protein assay, mass transfer coefficient, albumin

Affiliations:
Kupikowska-Stobba B. - IPPT PAN
Grzeczkowicz M. - other affiliation
Lewińska D. - Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
2.  Kupikowska-Stobba B., Lewińska D., Polymer microcapsules and microbeads as cell carriers for in vivo biomedical applications, Biomaterials Science, ISSN: 2047-4849, DOI: 10.1039/c9bm01337g, Vol.8, No.6, pp.1536-1574, 2020

Abstract:
Polymer microcarriers are being extensively explored as cell delivery vehicles in cell-based therapies and hybrid tissue and organ engineering. Spherical microcarriers are of particular interest due to easy fabrication and injectability. They include microbeads, composed of a porous matrix, and microcapsules, where matrix core is additionally covered with a semipermeable membrane. Microcarriers provide cell containment at implantation site and protect the cells from host immunoresponse, degradation and shear stress. Immobilized cells may be genetically altered to release a specific therapeutic product directly at the target site, eliminating side effects of systemic therapies. Cell microcarriers need to fulfil a number of extremely high standards regarding their biocompatibility, cytocompatibility, immunoisolating capacity, transport, mechanical and chemical properties. To obtain cell microcarriers of specified parameters, a wide variety of polymers, both natural and synthetic, and immobilization methods can be applied. Yet so far, only a few approaches based on cell-laden microcarriers have reached clinical trials. The main issue that still impedes progress of these systems towards clinical application is limited cell survival in vivo. Herein, we review polymer biomaterials and methods used for fabrication of cell microcarriers for in vivo biomedical applications. We describe their key limitations and modifications aiming at improvement of microcarrier in vivo performance. We also present the main applications of polymer cell microcarriers in regenerative medicine, pancreatic islet and hepatocyte transplantation and in the treatment of cancer. Lastly, we outline the main challenges in cell microimmobilization for biomedical purposes, the strategies to overcome these issues and potential future improvements in this area.

Affiliations:
Kupikowska-Stobba B. - other affiliation
Lewińska D. - Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
3.  Dorota Lewińska D., Grzeczkowicz M., Kupikowska-Stobba B., Influence of electric parameters on the alginate-polyethersulfone microcapsule structure, Desalination and Water Treatment, ISSN: 1944-3994, DOI: 10.5004/dwt.2017.11407, Vol.64, pp.400-408, 2017

Abstract:
Alginate-polyethersulfone microcapsules obtained with the one-stage three-nozzle, electrostatic technique designed by our team can successfully be used to encapsulate biologically active material such as functional proteins, microorganisms, bacteria, fungi and mammalian cells. The paper presents the results of studies on the influence of electric parameters on microcapsule size and internal structure. During the manufacturing process, three different liquids, i.e., aqueous sodium alginate solution, separating liquid and membrane-forming solution, were forced through a three-nozzle head, placed in an electrostatic field. After gelling in a gelling bath, the multi-layer drops appearing at the tip of the three-nozzle head formed microcapsules. The electrostatic field was applied through electric impulses with varied values of: voltage (U), frequency (f) and impulse duration (τ). The results indicated, that an increase of all examined electric process parameters resulted in a decrease in average microcapsule diameter and lower uniformity of batches in terms of size. Average membrane thickness (parameter B) did not change significantly, but along with the increase of all electrical parameters, a significant decrease of membrane thickness at the thickest part (parameter N) was observed. The microcapsules that were the most symmetric in regard to membrane thickness were obtained in the series with variable voltage (U).

Keywords:
alginate, microencapsulation of cells, alginate-polyethersulfone microcapsules, electrostatic technique, immobilization

Affiliations:
Dorota Lewińska D. - Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
Grzeczkowicz M. - other affiliation
Kupikowska-Stobba B. - other affiliation
4.  Przytulska M., Kulikowski Juliusz L., Lewińska D., Grzeczkowicz M., Kupikowska-Stobba B., Computer-aided image analysis for microcapsules’ quality assessment, Biocybernetics and Biomedical Engineering, ISSN: 0208-5216, DOI: 10.1016/j.bbe.2015.05.005, Vol.35, No.4, pp.342-350, 2015
5.  Kupikowska-Stobba B., Lewińska D., Grzeczkowicz M., Chemical method for retrieval of cells encapsulated in alginate-polyethersulfone microcapsules, Artificial Cells, Nanomedicine and Biotechnology, ISSN: 2169-141X, DOI: 10.3109/21691401.2013.800083, Vol.42, No.3, pp.151-160, 2014
6.  Chwojnowski A., Wojciechowski C., Lewińska D., Łukowska E., Nowak J., Kupikowska-Stobba B., Grzeczkowicz M., Studies on the structure of semipermeable membranes by means of SEM. Problems and potential sources of errors, Biocybernetics and Biomedical Engineering, ISSN: 0208-5216, DOI: 10.1016/s0208-5216(12)70032-3, Vol.32, No.1, pp.51-64, 2012
7.  Lewińska D., Chwojnowski A., Wojciechowski C., Kupikowska-Stobba B., Grzeczkowicz M., Weryński A., Electrostatic droplet generator with 3-coaxial-nozzle head for microencapsulation of living cells in hydrogel covered by synthetic polymer membranes, Separation Science and Technology, ISSN: 1520-5754, DOI: 10.1080/01496395.2011.617350, Vol.47, No.3, pp.463-469, 2012
8.  Kupikowska-Stobba B., Lewińska D., Dudziński K., Jankowska-Śliwińska J., Grzeczkowicz M., Wojciechowski C., Chwojnowski A., Influence of changes in composition of the membrane-forming solution on the structure of alginate-polyethersulfone microcapsules, Biocybernetics and Biomedical Engineering, ISSN: 0208-5216, Vol.29, No.3, pp.61-69, 2009

List of chapters in recent monographs
1. 
Lewińska D., Kupikowska-Stobba B., Grzeczkowicz M., Membranes and membrane processes in environmental protection. Monographs of Environmental Engineering Committee of Polish Academy of Sciences , rozdział: Influence of liquid flow rates on alginate-polyethersulfone microcapsule structure, Environmental Engineering Committee of Polish Academy of Sciences, 118, pp.367-380, 2014
2. 
Lewińska D., Grzeczkowicz M., Kupikowska-Stobba B., Membrany i procesy membranowe w ochronie środowiska. Monografie Komitetu Inżynierii Środowiska PAN , rozdział: Wpływ wielkości napięcia powierzchniowego cieczy żelującej na budowę membran w mikrokapsułkach glicerynowo-polieterosulfonowych, Komitet Inżynierii Środowiska PAN , 95, pp.93-106, 2012
3. 
Lewińska D., Kupikowska-Stobba B., Grzeczkowicz M., Wojciechowski C., Membrany i procesy membranowe w ochronie środowiska. Monografie Komitetu Inżynierii Środowiska PAN, rozdział: Wpływ parametrów elektrycznych na budowę mikrokapsułek glicerynowo-polieterosulfonowych, Komitet Inżynierii Środowiska PAN, 66, pp.9-22, 2010
4. 
Chwojnowski A., Wojciechowski C., Lewińska D., Kupikowska-Stobba B., Grzeczkowicz M., Dudziński K., Membrany i procesy membranowe w ochronie środowiska. Monografie Komitetu Inżynierii Środowiska PAN, rozdział: Badanie budowy membran półprzepuszczalnych za pomocą SEM. Problemy i potencjalne źródła błędów, Komitet Inżynierii Środowiska PAN, 65, pp.109-120, 2010
5. 
Kupikowska-Stobba B., Dudziński K., Chwojnowski A., Gutowska M., Sabalińska S., Lewińska D., Płończak M., Czubak J., Membrany i procesy membranowe w ochronie środowiska. Monografie Komitetu Inżynierii Środowiska PAN , rozdział: Wykorzystanie membran półprzepuszczalnych do hodowli chondrocytów króliczych, Komitet Inżynierii Środowiska PAN , 65, pp.341-352, 2010
6. 
Grzeczkowicz M., Kupikowska-Stobba B., Lewińska D., Membrany i procesy membranowe w ochronie środowiska. Monografie Komitetu Inżynierii Środowiska PAN , rozdział: Opracowanie preparatyki membran mikrokapsułek glicerynowo-polieterosulfonowych do badań mikroskopowych i komputerowej analizy obrazów, Komitet Inżynierii Środowiska PAN , 65, pp.185-197, 2010

Conference papers
1.  Grzeczkowicz M., Lewińska D., Kupikowska-Stobba B., Hydrogel-polyethersulfone microcapsules for cell storage, 138th International Centre of Biocybernetics seminar: Advances in Membrane and Adsorber Technology in Life Sciences, 2014-04-06/04-08, Warsaw (PL), pp.35-39, 2014

Conference abstracts
1.  Kupikowska-Stobba B., Grzeczkowicz M., Ładyżyński P., Lewińska D., Influence of surfactant concentration in gelling liquid on size and shape of polyethersulfone microcapsules produced from polymer solutions of different viscosities, XLV ESAO Congress, XLV Congress of the European Society for Artificial Organs, 2018-09-12/09-15, Madrid (ES), pp.600, 2018
2.  Lewińska D., Kupikowska-Stobba B., Grzeczkowicz M., Wpływ wielkości parametrów elektrycznych na budowę mikrokapsułek alginianowo-polieterosulfonowych, MEMPEP 2016, XI Krajowa Konferencja Membrany i procesy membranowe w ochronie środowiska, 2016-06-15/06-18, Zakopane (PL), pp.32, 2016
3.  Lewińska D., Kupikowska-Stobba B., Grzeczkowicz M., How to determine the concentration of cells encapsulated in alginate-polyethersulfone microcapsules, XLIII ESAO Congress, XLIII Congress of the European Society for Artificial Organs, 2016-09-14/09-17, Warsaw (PL), pp.345, 2016
4.  Lewińska D., Kupikowska-Stobba B., Grzeczkowicz M., Influence of liquid flow rates on alginate-polyethersulfone microcapsule structure, MEMPEP 2014, X Scientific Conference Membranes and Membrane Processes in Environmental Protection, 2014-06-06/06-08, Zakopane (PL), pp.380, 2014
5.  Lewińska D., Grzeczkowicz M., Kupikowska-Stobba B., Wpływ wielkości napięcia powierzchniowego cieczy żelującej na budowę membran w mikrokapsułkach glicerynowo-polieterosulfonowych, IX Krajowa Konferencja Membrany i procesy membranowe w ochronie środowiska, 2012-05-30/06-02, Zakopane (PL), pp.93, 2012
6.  Chwojnowski A., Wojciechowski C., Lewińska D., Kupikowska B., Grzeczkowicz M., Dudziński K., Badanie budowy membran półprzepuszczalnych za pomocą SEM. Problemy i potencjalne źródła błędów, MEMPEP 2010, VIII Krajowa Konferencja Membrany i procesy membranowe w ochronie środowiska, 2010-06-09/06-12, Zakopane (PL), pp.109, 2010
7.  Grzeczkowicz M., Kupikowska B., Lewińska D., Opracowanie preparatyki membran mikrokapsułek glicerynowo-polieterosulfonowych do badań mikroskopowych i komputerowej analizy obrazów, MEMPEP 2010, VIII Krajowa Konferencja Membrany i procesy membranowe w ochronie środowiska, 2010-06-09/06-12, Zakopane (PL), pp.185, 2010
8.  Lewińska D., Kupikowska B., Grzeczkowicz M., Wojciechowski C., Wpływ parametrów elektrycznych na budowę mikrokapsułek glicerynowo-polieterosulfonowych, MEMPEP 2010, VIII Krajowa Konferencja Membrany i procesy membranowe w ochronie środowiska, 2010-06-09/06-12, Zakopane (PL), pp.9, 2010
9.  Kupikowska B., Dudziński K., Chwojnowski A., Gutowska M., Sabalińska S., Lewińska D., Płończak M., Czubak J., Wykorzystanie membran półprzepuszczalnych do hodowli chondrocytów króliczych, MEMPEP 2010, VIII Krajowa Konferencja Membrany i procesy membranowe w ochronie środowiska, 2010-06-09/06-12, Zakopane (PL), pp.341, 2010
10.  Kupikowska B., Dudziński K., Jankowska-Śliwińska J., Grzeczkowicz M., Wojciechowski C., Lewińska D., Wpływ składu roztworu membranotwórczego na strukturę membran w mikrokapsułkach alginianowo-polieterosulfonowych, KBIB 2010, XVI Krajowa Konferencja Biocybernetyka i Inżynieria Biomedyczna, 2010-04-26/04-29, Warszawa (PL), pp.257, 2010
11.  Lewińska D., Kupikowska B., Grzeczkowicz M., Wojciechowski C., Wpływ przepływu cieczy na wielkość i strukturę mikrokapsułek polisacharydowo-polieterosulfonowych, KBIB 2010, XVI Krajowa Konferencja Biocybernetyka i Inżynieria Biomedyczna, 2010-04-26/04-29, Warszawa (PL), pp.144, 2010
12.  Grzeczkowicz M., Kupikowska B., Lewińska D., Elektrostatyczny generator kropel – precyzyjne narzędzie do mikroenkapsulacji komórek i substancji biologicznie aktywnych, KBIB 2010, XVI Krajowa Konferencja Biocybernetyka i Inżynieria Biomedyczna, 2010-04-26/04-29, Warszawa (PL), pp.258, 2010

Patents
Filing No./Date
Filing Publication
Autors
Title
Protection Area, Applicant Name
Patent Number
Date of Grant
pdf
402004
2012-12-11
BUP 13/2014
2014-06-23
Lewińska D., Kupikowska-Stobba B., Chwojnowski A., Grzeczkowicz M., Łukowska E.
Method for determining the concentration of cells
PL, Instytut Biocybernetyki i Inżynierii Biomedycznej PAN
223717
WUP 10/2016
2016-10-31



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